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  • NEBuffer 2.1

    This enzyme has transitioned to an improved new buffer system. Visit NEBCutSmart.com for further details.
    The new and current Double Digest Finder and current Activity/Performance Chart for the CutSmart buffer system are available. The previous version of the Double Digest Finder, as well as the previous Version of Activity/Performance Chart that use the former buffer system, are still available for your convenience.
    Catalog #SizeConcentrationPriceQtyAdd to Cart
    B7202S5 ml10X$18.00Add to Cart
    Buffer Products,
    Restriction Endonuclease Buffers & Diluents
    BioBrick® Assembly,
    DNA Nicking,
    Restriction Enzyme Digestion


    New England Biolabs provides a colorcoded 10X NEBuffer with each restriction endonuclease to ensure optimal (100%) activity. Most of our enzymes are supplied with one of four standard NEBuffers. Occasionally, an enzyme has specific buffer requirements not met by one of the four standard NEBuffers, in which case the enzyme is supplied with its own unique NEBuffer.

    Properties and Usage

    Storage Temperature


    1X Buffer Components

    50mM NaCl
    10mM Tris-HCl
    10mM MgCl2
    100μg/ml BSA
    pH 7.9@25°C

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Endonuclease Activity (Nicking):
      The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
    • Functional Test (Restriction Digest, Buffer):
      The buffer is tested for function by performing a restriction enzyme activity assay.
    • Non-Specific DNase Activity (16 hour):
      The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.
    • RNase Activity (Extended Digestion):
      The product is tested in a reaction containing a RNA substrate. After incubation for 16 hours greater than 90% of the substrate RNA remains intact as determined by gel electrophoresis.


    1. The pH of 10X NEBuffer 2.1 is betweenpH 7.8 and 8.0.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.


    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    1. What is supplied with the NEBuffer 2.1 pack?
    2. How should the NEBuffer be used?
    3. The buffer arrived thawed.  Are the buffer and the enzyme still active?
    4. I currently have an old tube of Restriction Enzyme - is it still active in the new buffer?
    5. You have replaced NEBuffer 1 with NEbuffer 1.1, NEBuffer 2 with NEBuffer 2.1, and NEBuffer 3 with 3.1. Where is NEBuffer 4.1?
    6. Why did you add BSA into all the restriction enzyme reaction buffers?
    7. Why did you remove DTT from your restriction enzyme buffers?
    8. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
    9. I don’t want to use BSA in my buffer. What are my options?